适合流式细胞仪分析的小麦细胞核提取方法比较  

Compare of extraction of wheat seedlings′ nucleus suitable for analysis by flow cytometry

作　　者：孙永星[1] 李素花[1] 魏小丽[1] 韩榕[1] 

机构地区：[1]山西师范大学生命科学学院,山西临汾041004

出　　处：《生物学杂志》2012年第3期88-91,共4页Journal of Biology

基　　金：国家自然科学基金资助项目(30671061);山西省自然科学基金资助项目(2008011059-1)

摘　　要：为制备供流式细胞仪分析的高纯度小麦细胞核悬液,以冬小麦"临优2018"为材料,分别采用酶解法和直接剪切法对其幼苗的细胞核进行提取,对所得到的细胞核在形态结构和数量等方面进行了分析和比较,根据其优缺点优化出最适合流式细胞仪分析的小麦幼苗细胞核的提取方法。结果显示:在直接剪切法所用的3种细胞核提取缓冲液中,MgSO4缓冲液的提取效果最好,细胞核形态及内部结构完整,且得到的细胞核量多;OttoⅠ对细胞核的提取效果显著,但是在加入OttoⅡ后细胞核破裂明显;Tris.MgCl2缓冲液提取的细胞核数量较少;酶解法制备的细胞核悬浮液中杂质较多,且需时较长。结果表明采用MgSO4提取缓冲液的直接剪切法是适合流式细胞仪对小麦幼苗DNA含量的分析。In order to prepare the high-purity wheat nucleus suspensions analyzed in the flow cytometry, winter wheat ＂linyou 2018＂ was used as research material for extraction of seedlings＇ nucleus with enzymatic and direct shear method. Nuclei＇s morphological structure and quantity was analyzed and compared. The most suitable extracting method of wheat seedlings＇ nuclei for the flow eytometry was obtained. The result showed that MgSO4 buffer was the best solution for nuclei extraction ; Otto I had a remarkable effect on the extrac- tion, but after with adding Otto 11 the nuclei broke obviously. Fewer nuclei were obtained with Tris · MgCl2 buffer. There are too many impurities in the nuclei suspension and long time with enzymatic method. The result indicated that MgSO4 extraction buffer of the direct shear method was suitable to analyze DNA content of wheat seedlings for flow eytometry.

关 键 词：小麦 流式细胞术 细胞核分离缓冲液 酶解法 

分 类 号：Q94-33[生物学—植物学] S512.1[农业科学—作物学]